Effects of HIV PIs on CNS endothelial cells in NeuroAIDS

HIV PI 对 NeuroAIDS 中中枢神经系统内皮细胞的影响

• 批准号: 6877966
• 负责人: Dianne Teresa LANGFORD
• 金额: $ 12.73万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6877966
• 关键词: AIDS /HIV neuropathy; AIDS therapy; HIV envelope protein gp120; HIV infections; P glycoprotein; SDS polyacrylamide gel electrophoresis; apoptosis; biological signal transduction; blood brain barrier; caveolins; central nervous system; clinical research; fibroblast growth factor; genetically modified animals; host organism interaction; immunocytochemistry; indinavir; laboratory mouse; mitogen activated protein kinase; nelfinavir; nitric oxide synthase; polymerase chain reaction; protease inhibitor; ritonavir; saquinavir; virus infection mechanism; western blottings

DESCRIPTION (provided by applicant): Long-term career goals are to: 1) develop a research program to investigate the cellular signaling alterations triggered by host-pathogen interactions relevant to neuro-psychiatric disorders such as HIV-associated Dementia (HAD) and 2) become an independent bio-medical scientist at an academic institution. Detailed career development and scientific plans addressing the potential contribution of highly active anti-retroviral therapy (HAART) to NeuroAIDS is proposed. Shortly after sero-conversion, HIV patients may be prescribed protease inhibitor (PI)-based HAART regimens. HAART has proven highly successful in suppressing systemic viral burden; however, the ATP-dependent efflux transport pump, P-glycoprotein (P-gp), imposes a serious constraint on PI treatment of NeuroAIDS. Because of viral rebound, resistance mutations and the potential interactions of PIs with HIV proteins and cerebral endothelial cells (CEC), the main objective of this proposal is to determine the effects of chronic PI exposure on the CEC's ability to respond to host-derived growth factors, such as fibroblast growth factor 2 (FGF2), that are generated as defense mechanisms against HIV proteins _resent in the blood stream at viral rebound. We hypothesize that long-term exposure to PIs such as saquinavir (SQV), indinavir (INV), nelfinavir (NFV) and/or ritonavir (RTV) will modify P-gp efflux-dependent expression and activity, thereby altering P-gp efflux-independent signaling pathways in the FGF2-mediated caveolin/ERK/NO systems. For this purpose, AIM I will determine the effects of chronic PI treatment on CEC fitness and signaling mediated by FGF2 via interactions with P-gp and caveolin. Four aspects of CEC fitness will be tested: 1) viability, 2) P-gp expression and activity, 3) P-gp-mediated caveolin and endothelial nitric oxide synthase (eNOS) signaling and nitric oxide (NO) production, 4) FGF2-mediated extracellular regulated kinase (ERK) signaling and angiogenic capacity. AIM II will address in vitro, how chronic exposure of CEC to PI disrupts P-gp/caveolin/FGF2-dependent protection from the HIV protein, gp120. Using MSR-gp120 and GFAPFGF2 transgenic mice, AIM III will investigate, in vivo, the long-term effects of chronic PI treatment in disrupting signaling at the blood-brain barrier (BBB) during interaction with HIV proteins. Understanding the mechanisms responsible for alterations in FGF2 signaling via P-gp/caveolae after chronic PI exposure is important for identifying factors that may contribute to the progression of neurological and neurobehavioral alterations associated with NeuroAIDS during viral rebound or at virologic failure.

描述（由申请人提供）：长期职业目标是： 1) 制定一项研究计划，调查与神经精神疾病相关的宿主-病原体相互作用所引发的细胞信号传导改变，例如 HIV 相关痴呆 (HAD) 和 2 ）成为学术机构的独立生物医学科学家。提出了详细的职业发展和科学计划，解决高效抗逆转录病毒疗法（HAART）对神经艾滋病的潜在贡献。血清转化后不久，HIV 患者可能会接受基于蛋白酶抑制剂 (PI) 的 HAART 治疗方案。 HAART 已被证明在抑制全身病毒负荷方面非常成功；然而，ATP依赖性外排转运泵P-糖蛋白(P-gp)对神经艾滋病的PI治疗施加了严重的限制。由于病毒反弹、耐药突变以及 PI 与 HIV 蛋白和脑内皮细胞 (CEC) 的潜在相互作用，本提案的主要目的是确定长期 PI 暴露对 CEC 响应宿主衍生生长的能力的影响因子，例如成纤维细胞生长因子 2 (FGF2)，是作为针对病毒反弹时血液中存在的 HIV 蛋白的防御机制而产生的。我们假设长期暴露于沙奎那韦 (SQV)、茚地那韦 (INV)、奈非那韦 (NFV) 和/或利托那韦 (RTV) 等 PI 将改变 P-gp 外排依赖性表达和活性，从而改变 P-gp 外排-FGF2介导的caveolin/ERK/NO系统中的独立信号通路。为此，AIM I 将确定慢性 PI 治疗对 CEC 适应性和 FGF2 通过与 P-gp 和 Caveolin 相互作用介导的信号传导的影响。将测试 CEC 适应性的四个方面：1) 活力，2) P-gp 表达和活性，3) P-gp 介导的小窝蛋白和内皮一氧化氮合酶 (eNOS) 信号传导和一氧化氮 (NO) 产生，4) FGF2 -介导的细胞外调节激酶（ERK）信号传导和血管生成能力。 AIM II 将在体外研究 CEC 长期暴露于 PI 如何破坏 HIV 蛋白 gp120 的 P-gp/caveolin/FGF2 依赖性保护。 AIM III 将使用 MSR-gp120 和 GFAPFGF2 转基因小鼠，在体内研究长期 PI 治疗在与 HIV 蛋白相互作用期间破坏血脑屏障 (BBB) 信号传导的长期影响。了解慢性 PI 暴露后通过 P-gp/caveolae 发生 FGF2 信号改变的机制对于识别可能导致病毒反弹或病毒学失败期间与 NeuroAIDS 相关的神经学和神经行为改变进展的因素非常重要。